Luminex 100 IS Developer Workbench Guide Version 2.3 User Manual
Page 66
Luminex 100 IS Developer Workbench Guide Version 2.3
x
MAP
Technology
60
PN 89-00002-00-084 Rev. B
5. Aspirate the supernatant and add 250 µL of the biotin-conjugated
molecule in Reaction Buffer. Vortex and sonicate for
approximately 20 seconds.
6. Dilute the biotin-conjugated molecule solution to a concentration
of 4 to 4000 nM.
7. Add 250 µL of the biotin-conjugated molecule solution to the
microsphere suspension and mix immediately by vortex.
8. Rotate the mixture for 30 minutes in the dark at room
temperature.
9. Centrifuge the xMAP microspheres at
≥ 8,000 × g for 1 to 2
minutes.
10. Aspirate the supernatant and resuspend the pelleted microspheres
in 500 µL of Blocking/Storage Buffer. Vortex.
11. Centrifuge the microspheres at
≥ 8,000 × g for 1 to 2 minutes.
12. Repeat steps 10 and 11 for a total of two washes with Blocking/
Storage Buffer.
13. Aspirate the supernatant and resuspend the microspheres in 250
to 1000 µL of Blocking/Storage Buffer. Vortex and sonicate for
about 20 seconds.
14. Enumerate the xMAP microsphere preparation.
15. Store the preparation at 2
o
C - 8°C. Protect it from light.
Technical Notes
1. The concentration of Biotin-conjugated molecules is a guide for
assay development.
For each specific biotin-conjugated molecule, you must
determine empirically the concentration required for optimal
reactivity.
2. The reaction volume can vary with the reagent volume. If the
reagent is limited, perform the reaction in a proportionately low
volume.
3. For best results, use microcentrifuge tubes from USA Scientific,
Inc., catalog number 1415-2500. To order, call 1 800 LAB-TIPS.